1. Field of the Invention
The present invention relates to certain 4-heteroaryl-3-heteroarylidenyl-2-indolinones compounds and their physiologically acceptable salts which modulate the activity of protein kinases (xe2x80x9cPKsxe2x80x9d). The compounds of the present invention are therefore useful in treating disorders related to abnormal PK activity. Pharmaceutical composition containing these compounds and methods of preparing these compounds are also described.
2. State of the Art
The following is offered as background information only and is not admitted to be prior art to the present invention.
PKs are enzymes that catalyze the phosphorylation of hydroxy groups on tyrosine, serine and threonine residues of proteins. The consequences of this seemingly simple activity are staggering; cell growth, differentiation and proliferation, i.e., virtually all aspects of cell life in one way or another depend on PK activity. Furthermore, abnormal PK activity has been related to a host of disorders, ranging from relatively non life threatening diseases such as psoriasis to extremely virulent diseases such as glioblastoma (brain cancer) (see U.S. Pat. No. 5,792,783 which is incorporated herein by reference in its entirety).
In view of the apparent link between PK-related cellular activities and wide variety of human disorders, a great deal of effort is being expended in an attempt to identify ways to modulate PK activity. Some of this effort has involved biomimetic approaches using large molecules patterned on those involved in the actual cellular processes (e.g., mutant ligands (U.S. Pat. No. 4,966,849); soluble receptors and antibodies (Published PCT Appl. WO 94/10202, Kendall and Thomas, Proc. Nat""l Acad. Sci., 90:10705-09 (1994), Kim, et al., Nature, 362:841-844 (1993)); RNA ligands (Jelinek, et al., Biochemistry, 33:10450-56); Takano, et al., Mol. Bio. Cell 4:358A (1993); Kinsella, et al., Exp. Cell Res. 199:56-62 (1992); Wright, et al., J. Cellular Phys., 152:448-57) and tyrosine kinase inhibitors (Published PCT Appls. WO 94/03427; 1 WO 92/21660; WO 91/15495; WO 94/14808; U.S. Pat. No. 5,330,992; Mariani, et al., Proc. Am. Assoc. Cancer Res., 35:2268 (1994)).
In addition to the above, attempts have been made to identify small molecules which act as PK inhibitors. For example, bis-monocylic, bicyclic and heterocyclic aryl compounds (Published PCT Appl. WO 92/20642), vinyleneazaindole derivatives (Published PCT Appl. WO 94/14808) and 1-cyclopropyl-4-pyridylquinolones (U.S. Pat. No. 5,330,992) have been described as tyrosine kinase inhibitors. Styryl compounds (U.S. Pat. No. 5,217,999), styryl-substituted pyridyl compounds (U.S. Pat. No. 5,302,606), quinazoline derivatives (EP App. No.0 566 266 A1), selenaindoles and selenides (Published PCT Appl. WO 94/03427), tricyclic polyhydroxylic compounds (Published PCT Appl. WO 92/21660), benzylphosphonic acid compounds (Published PCT Appl. WO 91/15495) and indolinone compounds (U.S. Pat. No. 5,792,783) have all been described as PTK inhibitors useful in the treatment of cancer. However these compounds have limited utility because of toxicity or poor bioavailability. Accordingly, there is a need for compounds that overcome these limitations. The compounds of the present invention fulfil this need.
In one aspect, this invention is directed to a compound of formula (I): 
wherein:
R1 and R2 are independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic, halo, xe2x80x94CX3, hydroxy, alkoxy, nitro, cyano, xe2x80x94C(O)R26, xe2x80x94C(O)OR26, R26C(O)Oxe2x80x94, xe2x80x94C(O)NR28R29, R26C(O)NR28xe2x80x94, xe2x80x94NR28R29,
xe2x80x94S(O)2R26, xe2x80x94S(O)2OR26, xe2x80x94S(O)2NR28R29, R26S(O)2NR28, X3CS(O)2xe2x80x94 and X3CS(O)2NR28xe2x80x94 where X is F, Cl, Br, or I;
Het is selected from the group consisting of: 
wherein:
A1, A2, A3, A4, and A5 are selected from the group consisting of carbon and nitrogen with the proviso that at least one and no more than two of A1, A2, A3, A4, and A5 are nitrogen;
R3, R4, R5, R6 and R7 are independently selected from the group consisting of hydrogen, alkyl, halo, hydroxy, alkoxy, X3Cxe2x80x94, nitro, cyano, xe2x80x94NR28R29, xe2x80x94C(O)OR and xe2x80x94C(O)NR28R29 where X is as defined above; it being understood that when A1, A2, A3, A4 or A5 is nitrogen, R3, R4, R5, R6 or R7, respectively, does not exist;
D is carbon or nitrogen;
R8, R9, R11 and R12 are independently selected from the group consisting of hydrogen, alkyl, hydroxy, alkoxy, halo, nitro, cyano and xe2x80x94NR28R29;
Z is selected from the group consisting of oxygen, sulfur, and xe2x80x94NR10;
R10 is selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, xe2x80x94C(O)R26, xe2x80x94C(S)R26, xe2x80x94C(O)OR26, xe2x80x94C(O)NR28R29, xe2x80x94C(S)NR28R29, xe2x80x94C(NH)NR28R29 and xe2x80x94S(O)2R26;
E1, E2, E3 and E4 are selected from the group consisting of carbon, nitrogen, oxygen and sulfur with the proviso that when D is carbon then at least one of E1, E2, E3 and E4 is other than carbon and that no more than one of E1, E2, E3 or E4 is oxygen or sulfur;
the dotted circle inside the five-member ring contain D, E1, E2, E3 and E4 ring means that the ring system is aromatic;
R13, R14, R15 and R16 are independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, mercapto, thioalkoxy, halo, nitro, cyano, xe2x80x94C(O)R26, xe2x80x94C(O)OR26, xe2x80x94C(O)NR28R29 and xe2x80x94NR28R29, it being understood that, when one of E1, E2, E3 or E4 is sulfur or oxygen and any of the others is nitrogen, there is no R group bonded to any of those nitrogens, it also being understood that, when two or three of E1, E2, E3 or E4 are nitrogen, there is an R group bonded to one of the nitrogens and that R group is selected from the group consisting of hydrogen and alkyl, there being no R group bonded to any of the other nitrogens;
Q is selected from the group consisting of: 
where:
G1, G2, G3, G4 and G5 are selected from the group consisting of carbon and nitrogen with the proviso that no more than two of G1, G2, G3, G4 and G5 are nitrogen;
R17, R18, R19, R20 and R21 are independently selected from the group consisting of hydrogen, alkyl, hydroxy, alkoxy, halo,
xe2x80x94NR28R29, xe2x80x94(CH2)nC(O)R26, xe2x80x94(CH2)nC(O)OR26 and xe2x80x94(CH2)nC(O)NR28R29,
xe2x80x94(CH2)nNR28R29, xe2x80x94(CH2)nS(O)2R26 and xe2x80x94(CH2)nS(O)2NR28R29;
J1 is selected from the group consisting of nitrogen, oxygen and sulfur such that when J1 is nitrogen, R22 is selected from the group consisting of hydrogen, alkyl and xe2x80x94C(O)R26; and
when J1 is oxygen or sulfur, R22 does not exist;
J2, J3 and J4 are selected from the group consisting of carbon and nitrogen;
R23, R24 and R25 are independently selected from the group consisting of hydrogen, alkyl, aryl optionally substituted with one or more groups independently selected from the group consisting of hydroxy, unsubstituted lower alkoxy and halo, halo,
xe2x80x94(CH2)nC(O)R26, xe2x80x94(CH2)nC(O)OR26 and xe2x80x94(CH2)nC(O)NR28R29, xe2x80x94(CH2)nNR28R29,
xe2x80x94(CH2)nS(O)2R26, xe2x80x94(CH2)nS(O)2NR28R29, xe2x80x94(CH2)nOR26, xe2x80x94O(CH2)nNR28R29 and xe2x80x94C(O)NH (CH2)nNR28R29;
n is 0, 1, 2, or 3;
R23 and R24 or R24 and R25 may combine to form a group selected from the group consisting of xe2x80x94CH2CH2CH2CH2xe2x80x94, xe2x80x94CHxe2x95x90CR33xe2x80x94CR34xe2x95x90CHxe2x80x94 and
xe2x80x94C(O)Y(CH2)2xe2x80x94 and group wherein Y is selected from the group consisting of oxygen, sulfur and xe2x80x94N(R27)xe2x80x94 and R33 and R34 are selected from the group consisting of hydrogen, xe2x80x94(CH2)nNR28R29 and xe2x80x94O(CH2)nNR28R29 where, when one of R33 or R34 is xe2x80x94(CH2)nNR28R29 or
xe2x80x94O(CH2)nNR28R29, the other is hydrogen;
it being understood that, when J2, J3 or J4 is nitrogen, R23, R24 or R25, respectively, does not exist;
R26 is selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl and heteroaryl;
R27 is selected from the group consisting of hydrogen and alkyl;
R28 and R29 are independently selected from the group consisting of hydrogen, alkyl, aryl, heteroaryl, xe2x80x94(CH2)naryl, xe2x80x94(CH2)nheteroaryl and xe2x80x94C(O)R26, or, combined, R28 and R29 may form a group selected from the group consisting of xe2x80x94(CH2)5xe2x80x94, xe2x80x94(CH2)2O(CH2)2xe2x80x94, xe2x80x94(CH2)2NR3 (CH2)2xe2x80x94 and xe2x80x94(CH)3C(O)xe2x80x94 wherein R30 is selected from the group consisting of hydrogen, alkyl, xe2x80x94C(O)R26, xe2x80x94S(O)2R26, xe2x80x94S(O)3R26, xe2x80x94S(O)2NR31R32, xe2x80x94C(O)NHNR31R32, xe2x80x94C(O)NR31R32, xe2x80x94C(S)NR31R32 and xe2x80x94C(O)OR26 where R31 and R32 are independently selected from the group consisting of hydrogen, unsubstituted lower alkyl and aryl optionally substituted with one or more groups independently selected from the group consisting of halo and unsubstituted lower alkoxy; or a pharmaceutically acceptable salt thereof; provided that: the compound of formula (I) is not:
(Z)-1,3-dihydro-3-[(1H-pyrrol-2-yl)methylene]-4-(2-thiophenyl)-2H-indol-2-one; and
Z)-1,3-dihydro-4-(2,4-dimethoxy-6-pyrimidinyl)-3-[(1H-pyrrol-2-yl)methylene]-2H-indol-2-one.
In a second aspect, this invention relates to a method for the modulation of the catalytic activity of a PK by contacting a PK with a compound of this invention or a pharmaceutically acceptable salt thereof. The modulation of the catalytic activity of PKs using a compound of this invention may be carried out in vitro or in vivo.
The protein kinase whose catalytic activity is being modulated by a compound of this invention is selected from the group consisting of receptor protein tyrosine kinases, cellular (or non-receptor) tyrosine kinases and serine-threonine kinases.
Preferably, the receptor protein kinase whose catalytic activity is modulated by a compound of this invention is selected from the group consisting of EGF, HER2,HER3,HER4, IR, IGF-1R, IRR, PDGFRxcex1, PDGFRxcex2, CSFIR, C-Kit, C-fms, Flk-1R, Flk4, KDR/Flk-1, Flt-1, FGFR-1R, FGFR-2R, FGFR-3R and FGFR-4R. The cellular tyrosine kinase whose catalytic activity is modulated by a compound of this invention is selected from the group consisting of Src, Frk, Btk, Csk, Abl, ZAP70, Fes/Fps, Fak, Jak, Ack, Yes, Fyn, Lyn, Lck, Blk, Hck, Fgr and Yrk. The serine-threonine protein kinase whose catalytic activity is modulated by a compound of this invention is selected from the group consisting of CDK2 and Raf.
In a third aspect, this invention is directed to a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient. Such pharmaceutical composition may contain both carriers and excipients as well as other components generally known to those skilled in the formulation of pharmaceutical compositions.
In a fourth aspect, this invention is directed to a method for treating or preventing a protein kinase related disorder in an organism which method comprises administering to said organism a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.
The above-referenced protein kinase related disorders are those mediated by receptor protein tyrosine kinases, non-receptor or cellular tyrosine kinases, and serine-threonine kinases.
Preferably, the protein kinase related disorders are those mediated by EGFR, a PDGFR, IGFR, flk (VEGFR), CDK2, Met kinase, and Src kinase.
More preferably, the disorders are cancer selected from the group consisting of squamous cell carcinoma, sarcomas such as Kaposi""s sarcoma, astrocytoma, glioblastoma, lung cancer, bladder cancer, colorectal cancer, gastrointestinal cancer, head and neck cancer, melanoma, ovarian cancer, prostate cancer, breast cancer, small-cell lung cancer, glioma, colorectal cancer, genitourinary cancer and gastrointestinal cancer; diabetic retinopathy, a hyperproliferation disorder, von Hippel-Lindau disease, restenosis, fibrosis, psoriasis, inflammatory disorders such as rheumatoid arthritis, osteoarthritis, immunological disorders such as autoimmune diseases, cardiovasular disorders such as atherosclerosis and angiogenesis related disorders.
In a fifth aspect, this invention is directed to a use of a compound of formula (I) as a reference compound in an assay in order to identify new compounds (test compounds) that modulate protein kinase activity which method comprises contacting cells expressing said protein kinase with a test compound or a compound of formula (I) and then monitoring said cells for an effect.
The above-referenced effect is selected from a change or an absence of change in a cell phenotype, a change or absence of change in the catalytic activity of said protein kinase or a change or absence of change in the interaction of said protein kinase with a natural binding partner.
Definitions:
The following terms used in the claims and the specification have the meanings given below. Other terms have their art recognized meaning.
The term xe2x80x9calkylxe2x80x9d refers to a saturated aliphatic hydrocarbon including straight chain and branched chain groups. Preferably, the alkyl group has 1 to 20 carbon atoms (whenever a numerical range; e.g. xe2x80x9c1-20xe2x80x9d, is stated herein, it means that the group, in this case the alkyl group, may contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 20 carbon atoms). More preferably, it is a medium size alkyl radical having 1 to 10 carbon atoms e.g., methyl, ethyl, propyl, 2-propyl, n-butyl, iso-butyl, tert-butyl, pentyl, and the like. Most preferably, it is lower alkyl having 1 to 4 carbon atoms e.g., methyl, ethyl, propyl, 2-propyl, n-butyl, iso-butyl, or tert-butyl, and the like.
The alkyl group may be substituted or unsubstituted. When substituted, the substituent group(s) is preferably one, two, or three, more preferably one or two groups, independently selected from the group consisting of halo, hydroxy, unsubstituted lower alkoxy, mercapto, (unsubstituted lower alkyl)thio, cyano, nitro,
xe2x80x94C(O)R33, xe2x80x94C(S)R33, xe2x80x94OC(O)NR34R35, R33OC(O)NR34xe2x80x94, xe2x80x94C(S)NR34R35, R33OC(S)NR34xe2x80x94, xe2x80x94C(O)NR34R35, R33C(O)NR34xe2x80x94, R33S(O)2NR34xe2x80x94, xe2x80x94S(O)2NR34R35 R33 S(O)xe2x80x94, R33S(O)2xe2x80x94, xe2x80x94C(O)OR33, R33C(O)Oxe2x80x94, xe2x80x94NR34R3, aryl optionally substituted with one or more, more preferably one, two, or three groups independently selected from the group consisting of halo, hydroxy and unsubstituted lower alkoxy, aryloxy optionally substituted with one or more, more preferably one, two, or three groups independently selected from the group consisting of halo, hydroxy and unsubstituted lower alkoxy, arylthio optionally substituted with one or more, more preferably one, two, or three groups independently selected from the group consisting of halo, hydroxy and unsubstituted lower alkoxy, 6-member heteroaryl having from 1 to 3 nitrogen atoms in the ring, the carbons in the ring being optionally substituted with one or more, more preferably one or two groups independently selected from the group consisting of halo, hydroxy and unsubstituted lower alkoxy, 5-member heteroaryl having from 1 to 3 heteroatoms in the ring selected from the group consisting of nitrogen, oxygen and sulfur, the carbon atoms of the group being optionally substituted with one or more, more preferably one or two groups independently selected from the group consisting of halo, hydroxy and unsubstituted lower alkoxy groups and a 5- or 6-member heteroalicyclic group having from 1 to 3 heteroatoms in the ring selected from the group consisting of nitrogen, oxygen and sulfur, the carbon atoms in the group being optionally substituted with one or more, more preferably one or two groups independently selected from the group consisting of halo, hydroxy and unsubstituted lower alkoxy groups, wherein R33 is selected from the group consisting of hydrogen, unsubstituted lower alkyl and aryl optionally substituted with one or more, more preferably one, two, or three groups independently selected from the group consisting of halo and unsubstituted lower alkoxy and R34 and R35 are independently selected from the group consisting of hydrogen, unsubstituted lower alkyl, xe2x80x94C(O)R33, aryl optionally substituted with one, two, or three groups independently selected from the group consisting of halo and unsubstituted lower alkoxy and heteroaryl optionally substituted with one, two, or three groups independently selected from the group consisting of halo and unsubstituted lower alkoxy.
A xe2x80x9ccycloalkylxe2x80x9d group refers to a 3 to 8 member all-carbon monocyclic ring, an all-carbon 5-member/6-member or 6-member/6-member fused bicyclic ring or a multicyclic fused ring (a xe2x80x9cfusedxe2x80x9d ring system means that each ring in the system shares an adjacent pair of carbon atoms with each other ring in the system) group wherein one or more of the rings may contain one or more double bonds but none of the rings has a completely conjugated pi-electron system. Examples, without limitation, of cycloalkyl groups are cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, cyclohexadiene, adamantane, cycloheptane and, cycloheptatriene. A cycloalkyl group may be substituted or unsubstituted. When substituted, the substituent group(s) is preferably one or more independently selected from the group consisting of halo, hydroxy, unsubstituted lower alkoxy, mercapto, (unsubstituted lower alkyl)thio, cyano, nitro, xe2x80x94C(O)R33xe2x80x94C(S)R33, xe2x80x94OC(O)NR34R35, R33OC(O)NR34xe2x80x94, xe2x80x94OC(S)NR34R35, R33OC(S)NR34xe2x80x94, xe2x80x94C(O)NR34R35, R33C(O)NR34xe2x80x94, R33S(O)2NR34xe2x80x94, xe2x80x94S(O)2NR34R35, R33S(O)xe2x80x94, R33S(O)2xe2x80x94, xe2x80x94C(O)OR33, R33C(O)Oxe2x80x94, xe2x80x94NR34R35, aryl optionally substituted with one, two or three groups independently selected from the group consisting of halo, hydroxy and unsubstituted lower alkoxy, aryloxy optionally substituted with with one, two or three groups independently selected from the group consisting of halo, hydroxy and unsubstituted lower alkoxy, arylthio optionally substituted with with one, two or three groups independently selected from the group consisting of halo, hydroxy and unsubstituted lower alkoxy, 6-member heteroaryl having from 1 to 3 nitrogen atoms in the ring, the carbons in the ring being optionally substituted with with one, two or three groups independently selected from the group consisting of halo, hydroxy and unsubstituted lower alkoxy, 5-member heteroaryl having from 1 to 3 heteroatoms in the ring selected from the group consisting of nitrogen, oxygen and sulfur, the carbon atoms of the group being optionally substituted with one or two groups independently selected from the group consisting of halo, hydroxy and unsubstituted lower alkoxy groups and a 5- or 6-member heteroalicyclic group having from 1 to 3 heteroatoms in the ring selected from the group consisting of nitrogen, oxygen and sulfur, the carbon atoms in the group being optionally substituted with with one, two or three groups independently selected from the group consisting of halo, hydroxy and unsubstituted lower alkoxy groups, wherein R33 is selected from the group consisting of hydrogen, unsubstituted lower alkyl and aryl optionally substituted with one, two or three groups independently selected from the group consisting of halo and unsubstituted lower alkoxy and R34 and R35 are independently selected from the group consisting of hydrogen, unsubstituted lower alkyl, xe2x80x94C(O)R33, aryl optionally substituted with with one, two or three groups independently selected from the group consisting of halo and unsubstituted lower alkoxy and heteroaryl optionally substituted with with one, two or three groups independently selected from the group consisting of halo and unsubstituted lower alkoxy.
An xe2x80x9calkenylxe2x80x9d group refers to an alkyl group, as defined herein, consisting of at least two carbon atoms and at least one carbonxe2x80x94carbon double bond e.g., ethenyl, propenyl, butenyl, and the like.
An xe2x80x9calkynylxe2x80x9d group refers to an alkyl group, as defined herein, consisting of at least two carbon atoms and at least one carbonxe2x80x94carbon triple bond e.g., ethynyl, propynyl, and the like.
An xe2x80x9carylxe2x80x9d group refers to an all-carbon monocyclic or fused-ring polycyclic (i.e., rings which share adjacent pairs of carbon atoms) groups of 6 to 12 ring atoms and having a completely conjugated pi-electron system. Examples, without limitation, of aryl groups are phenyl, naphthalenyl and anthracenyl. The aryl group may be substituted or unsubstituted. When substituted, the substituted group(s) is preferably one or more, more preferably one, two, or three, independently selected from the group consisting of unsubstituted lower alkyl, X3Cxe2x80x94, halo, hydroxy, unsubstituted lower alkoxy, mercapto, (unsubstituted lower alkyl)thio, cyano, nitro, xe2x80x94C(O) R33, xe2x80x94C(S)R33, xe2x80x94C(O)NR3R35, R33OC(O)NR34xe2x80x94, xe2x80x94OC(S)NR34R35, R33OC(S)NR34xe2x80x94, xe2x80x94C(O)NR34R35, R33C(O)NR34xe2x80x94, R33S(O)2NR34xe2x80x94, xe2x80x94S(O)2NR34R35, R33S(O)xe2x80x94, R33S(O)2xe2x80x94, xe2x80x94C(O)OR33, R33C(O)Oxe2x80x94 and xe2x80x94NR34R35 with R33, R34 and R35 as defined above.
As used herein, a xe2x80x9cheteroarylxe2x80x9d group refers to a monocyclic or fused aromatic ring (i.e., rings which share an adjacent pair of atoms) containing 5 to 10 ring atoms wherein one, two, three or four ring atoms are independently selected from the group consisting of nitrogen, oxygen and sulfur, the rest being carbon. Examples, without limitation, of heteroaryl groups are pyrrole, furan, thiophene, imidazole, oxazole, isoxazole, thiazole, pyrazole, pyridine, pyrimidine, pyrazine, pyridazine, quinoline, isoquinoline, purine and carbazole. With regard to the five-member heteroaryl groups containing two or more nitrogens and no other hetero atoms in the ring, such as imidazole and triazole, one of the nitrogens in the ring may be bonded to an R group while the others may not. This gives rise to isomeric structures such as those shown below for dimethylimidazoles and dimethyltriazoles: 
All such isomers are within the scope of this invention. A heteroaryl group may be substituted or unsubstituted. When substituted, the substituted group(s) is preferably one or more, more preferably one or two groups independently selected from the group consisting of unsubstituted lower alkyl, X3Cxe2x80x94, halo, hydroxy, unsubstituted lower alkoxy, mercapto, (unsubstituted lower alkyl)thio, cyano, nitro, xe2x80x94C(O)R33, xe2x80x94C(S)R33, xe2x80x94OC(O)NR34R35, R33OC(O)NR34xe2x80x94, xe2x80x94OC(S)NR34R35, R33OC(S)NR34xe2x80x94, xe2x80x94C(O)NR34R35, R33C(O)NR34xe2x80x94, R33S(O)2NR34xe2x80x94, xe2x80x94S(O)2NR34R35, R33S(O)xe2x80x94, R33S(O)2xe2x80x94, xe2x80x94C(O)OR33, R33C(O)Oxe2x80x94 and xe2x80x94NR34R35 with R33, R34 and R35 as defined above.
A xe2x80x9cheteroalicyclicxe2x80x9d group refers to a monocyclic or fused ring of 5 to 10 ring atoms wherein one, two, or three ring atoms are independently selected from the group consisting of nitrogen, oxygen and sulfur, the rest being carbon. The rings may also have one or more double bonds. However, the rings do not have a completely conjugated pi-electron system. The heteroalicyclic ring may be substituted or unsubstituted. When substituted, the substituted group(s) is preferably one or more, more preferably one or two groups independently selected from the group consisting of unsubstituted lower alkyl, X3Cxe2x80x94, halo, hydroxy, unsubstituted lower alkoxy, mercapto, (unsubstituted lower alkyl)thio, cyano, nitro, xe2x80x94C(O)R33, xe2x80x94C(S)R33, xe2x80x94OC(O)NR34R35, R33C(O)NR3xe2x80x94, xe2x80x94OC(S)NR34R35, R33OC(S)NR34xe2x80x94, xe2x80x94C(O)NR34R35, R33C(O)NR xe2x80x94, R33S(O)2NR34xe2x80x94, xe2x80x94S(O)2NR34R35, R33S(O)xe2x80x94, R33S(O)2xe2x80x94, xe2x80x94C(O)OR33, R33C(O)Oxe2x80x94 and xe2x80x94NR34R35 with R33, R34 and R35 as defined above.
A xe2x80x9chydroxyxe2x80x9d group refers to an xe2x80x94OH group.
An xe2x80x9calkoxyxe2x80x9d group refers to both an xe2x80x94O-(unsubstituted alkyl) and an xe2x80x94O-(unsubstituted cycloalkyl) group.
An xe2x80x9caryloxyxe2x80x9d group refers to both an xe2x80x94O-aryl and an xe2x80x94O-heteroaryl group, as defined herein.
A xe2x80x9cmercaptoxe2x80x9d group refers to an xe2x80x94SH group.
A xe2x80x9calkylthioxe2x80x9d group refers to both an xe2x80x94S(unsubstituted alkyl) and an xe2x80x94S(unsubstituted cycloalkyl) group.
A xe2x80x9cthioalkoxyxe2x80x9d group refers to both an S-alkyl and an xe2x80x94S-cycloalkyl group, as defined herein.
A xe2x80x9carylthioxe2x80x9d group refers to both an xe2x80x94S(aryl) and an xe2x80x94S(heteroaryl group), as defined herein.
A xe2x80x9chaloxe2x80x9d group refers to fluorine, chlorine, bromine or iodine.
A xe2x80x9ccyanoxe2x80x9d group refers to a xe2x80x94Cxe2x89xa1N group.
A xe2x80x9cnitroxe2x80x9d group refers to a xe2x80x94NO2 group.
xe2x80x9cHeteroarylxe2x80x9d refers to both heteroaryl groups, defined elsewhere herein and exemplified, without limitation, by the compounds of Group I, below, and heteroalicyclic groups, likewise defined elsewhere herein and, again without limitation, exemplified by the compounds of Group II, below: 
As used herein xe2x80x9cheteroarylidenylxe2x80x9d refers to a group having the following structure, wherein Q is a heteroaryl group, as defined above. 
The terms xe2x80x9c2-oxindole,xe2x80x9d xe2x80x9c2-indolinone,xe2x80x9d and xe2x80x9cindolin-2-onexe2x80x9d are used interchangeable to refer to a group having the following structure. The 3 and 4 positions, wherein compounds of this invention are substituted with a heteroarylidenyl or a heteroaryl group, respectively, are marked: 
As used herein, the term xe2x80x9ccombined,xe2x80x9d when referring to two R groups bonded to adjacent carbon atoms, means that the atoms shown as comprising the xe2x80x9ccombinedxe2x80x9d structure form a bridge from the carbon to which one of the R groups is bonded to the carbon atom to which the other R group is bonded.
As used herein, xe2x80x9cPKxe2x80x9d refers to receptor protein tyrosine kinase (RTKs), non-receptor or xe2x80x9ccellularxe2x80x9d tyrosine kinase (CTKs) and serine-threonine kinases (STKs).
The term xe2x80x9cmethodxe2x80x9d refers to manners, means, techniques and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques and procedures either known to, or readily developed from known manners, means, techniques and procedures by, practitioners of the chemical, pharmaceutical, biological, biochemical and medical arts.
As used herein, the term xe2x80x9cmodulationxe2x80x9d or xe2x80x9cmodulatingxe2x80x9d refers to the alteration of the catalytic activity of RTKs, CTKs and STKs. In particular, modulating refers to the activation or inhibition of the catalytic activity of RTKs, CTKs and STKs, preferably the activation or inhibition of the catalytic activity of RTKs, CTKs and STKs, depending on the concentration of the compound or salt to which the RTK, CTK or STK is exposed or, more preferably, the inhibition of the catalytic activity of RTKs, CTKs and STKs.
The term xe2x80x9ccatalytic activityxe2x80x9d as used herein refers to the rate of phosphorylation of tyrosine under the influence, direct or indirect, of RTKs and/or CTKs or the phosphorylation of serine and threonine under the influence, direct or indirect, of STKs.
The term xe2x80x9ccontactingxe2x80x9d as used herein refers to bringing a compound of this invention and a target PK together in such a manner that the compound can affect the catalytic activity of the PK, either directly, i.e., by interacting with the kinase itself, or indirectly, i.e., by interacting with another molecule on which the catalytic activity of the kinase is dependent. Such xe2x80x9ccontactingxe2x80x9d can be accomplished xe2x80x9cin vitro,xe2x80x9d i.e., in a test tube, a petri dish or the like. In a test tube, contacting may involve only a compound and a PK of interest or it may involve whole cells. Cells may also be maintained or grown in cell culture dishes and contacted with a compound in that environment. In this context, the ability of a particular compound to affect a PK related disorder, i.e., the IC50 of the compound, defined below, can be determined before use of the compounds in vivo with more complex living organisms is attempted. For cells outside the organism, multiple methods exist, and are well-known to those skilled in the art, to get the PKs in contact with the compounds including, but not limited to, direct cell microinjection and numerous transmembrane carrier techniques.
A xe2x80x9cpharmaceutical compositionxe2x80x9d refers to a mixture of one or more of the compounds described herein, or physiologically/pharmaceutically acceptable salts or prodrugs thereof, with other chemical components, such as physiologically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration of a compound to an organism.
A xe2x80x9cprodrugxe2x80x9d refers to an agent which is converted into the parent drug in vivo. Prodrugs are often useful because, in some situations, they may be easier to administer than the parent drug. They may, for instance, be bioavailable by oral administration whereas the parent drug is not. The prodrug may also have improved solubility in pharmaceutical compositions over the parent drug. An example, without limitation, of a prodrug would be a compound of the present invention which is administered as an ester (the xe2x80x9cprodrugxe2x80x9d) to facilitate transmittal across a cell membrane where water solubility is detrimental to mobility but then is metabolically hydrolyzed to the carboxylic acid, the active entity, once inside the cell where water solubility is beneficial.
A further example of a prodrug might be a short polypeptide, for example, without limitation, a 2-10 amino acid polypeptide, bonded through a terminal amino group to a carboxy group of a compound of this invention wherein the polypeptide is hydrolyzed or metabolized in vivo to release the active molecule.
As used herein, a xe2x80x9cphysiologically/pharmaceutically acceptable carrierxe2x80x9d refers to a carrier or diluent that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound.
An xe2x80x9cexcipientxe2x80x9d refers to an inert substance added to a pharmaceutical composition to further facilitate administration of a compound. Examples, without limitation, of excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils and polyethylene glycols.
xe2x80x9cIn vitroxe2x80x9d refers to procedures performed in an artificial environment such as, e.g., without limitation, in a test tube or culture medium.
As used herein, xe2x80x9cin vivoxe2x80x9d refers to procedures performed within a living organism such as, without limitation, a mouse, rat or rabbit.
As used herein, xe2x80x9cPK related disorder,xe2x80x9d xe2x80x9cPK driven disorder,xe2x80x9d and xe2x80x9cabnormal PK activityxe2x80x9d all refer to a condition characterized by inappropriate, i.e., under or, more commonly, over, PK catalytic activity, where the particular PK can be an RTK, a CTK or an STK. Inappropriate catalytic activity can arise as the result of either: (1) PK expression in cells which normally do not express PKs, (2) increased PK expression leading to unwanted cell proliferation, differentiation and/or growth, or, (3) decreased PK expression leading to unwanted reductions in cell proliferation, differentiation and/or growth. Over-activity of a PK refers to either amplification of the gene encoding a particular PK or production of a level of PK activity which can correlate with a cell proliferation, differentiation and/or growth disorder (that is, as the level of the PK increases, the severity of one or more of the symptoms of the cellular disorder increases). Under-activity is, of course, the converse, wherein the severity of one or more symptoms of a cellular disorder increase as the level of the PK activity decreases.
As used herein, the terms xe2x80x9cpreventxe2x80x9d, xe2x80x9cpreventingxe2x80x9d and xe2x80x9cpreventionxe2x80x9d refer to a method for barring an organism from acquiring a PK related disorder in the first place.
As used herein, the terms xe2x80x9ctreatxe2x80x9d, xe2x80x9ctreatingxe2x80x9d and xe2x80x9ctreatmentxe2x80x9d refer to a method of alleviating or abrogating a PK mediated cellular disorder and/or its attendant symptoms. With regard particularly to cancer, these terms simply mean that the life expectancy of an individual affected with a cancer will be increased or that one or more of the symptoms of the disease will be reduced.
The term xe2x80x9corganismxe2x80x9d refers to any living entity comprised of at least one cell. A living organism can be as simple as, for example, a single eukariotic cell or as complex as a mammal, such as a cat, dog, human being, etc.
The term xe2x80x9ctherapeutically effective amountxe2x80x9d as used herein refers to that amount of the compound being administered which will relieve to some extent one or more of the symptoms of the disorder being treated. In reference to the treatment of cancer, a therapeutically effective amount refers to that amount which has the effect of (1) reducing the size of the tumor, (2) inhibiting (that is, slowing to some extent, preferably stopping) tumor metastasis, (3) inhibiting to some extent (that is, slowing to some extent, preferably stopping) tumor growth, and/or, (4) relieving to some extent (or, preferably, eliminating) one or more symptoms associated with the cancer.
By xe2x80x9cmonitoringxe2x80x9d is meant observing or detecting the effect of contacting a compound with a cell expressing a particular PK. The observed or detected effect can be a change in cell phenotype, in the catalytic activity of a PK or a change in the interaction of a PK with a natural binding partner. Techniques for observing or detecting such effects are well-known in the art.
xe2x80x9cCell phenotypexe2x80x9d refers to the outward appearance of a cell or tissue or the biological function of the cell or tissue. Examples, without limitation, of a cell phenotype are cell size, cell growth, cell proliferation, cell differentiation, cell survival, apoptosis, and nutrient uptake and use. Such phenotypic characteristics are measurable by techniques well-known in the art.
A xe2x80x9cnatural binding partnerxe2x80x9d refers to a polypeptide that binds to a particular PK in a cell. Natural binding partners can play a role in propagating a signal in a PK-mediated signal transduction process. A change in the interaction of the natural binding partner with the PK can manifest itself as an increased or decreased concentration of the PK/natural binding partner complex and, as a result, in an observable change in the ability of the PK to mediate signal transduction.
At present certain compounds of formula (I) are more preferred. Some such preferred embodiments are disclosed below:
(i) A preferred group of compounds is that wherein Het is: 
wherein:
A1 or A2 or A3 or A2 and A4 are nitrogen and the other A""s are carbon and the R groups on the A""s that are carbon are selected from the group consisting of hydrogen, xe2x80x94NH2 and xe2x80x94C(O)OR26, R26 being selected from the group consisting of hydrogen and unsubstituted lower alkyl. More preferably Het is 2-, 3-, or 4-pyridyl or 2-, 4-, or 5-pyrimidinyl optionally substituted with an amino or xe2x80x94COOH group. Most preferably 4-pyridyl.
(ii) Another presently preferred embodiment of this invention is that wherein Het is: 
wherein:
D is nitrogen or carbon, preferably carbon;
R8, R9, R11 and R12 are hydrogen; and
Z is xe2x80x94NR10 where R10 is selected from the group consisting of hydrogen, xe2x80x94C(O)R26, xe2x80x94C(O)OR26, xe2x80x94C(O)NR28R29, xe2x80x94C(S)NR28R29, xe2x80x94C(NH)NR28R29 and xe2x80x94S(O)2R26 where R26, R28, and R29 are as defined in the Summary of the invention Preferably Het is piperidin-4-yl, piperazin-4-yl, or 4-methylpiperazin-1-yl.
(iii) Another presently preferred aspect of this invention is that wherein Het is: 
wherein:
D is carbon, E1 is sulfur, E4 is nitrogen, E2 and E3 are carbon, R13 and R16 do not exist and R14 and R15 are hydrogen or E2 is nitrogen, E4 is sulfur, E1 and E3 are carbon, R13 is hydrogen, R14 and R16 do not exist and R15 is xe2x80x94NR28R29 or E2 and E3 are nitrogen, E1 and E4 are carbon, R13 and R16 are hydrogen and R14 and R15 do not exist. Preferably, Het is thiazol-2-yl.
(iv) Another preferred group of compounds is that wherein Q is: 
wherein:
J1 is nitrogen and J2, J3 and J4 are carbon.
Within this group a more preferred group of compounds is that wherein R22 is hydrogen.
Within the more preferred group, an even more preferred group of compounds is that wherein:
R23 is selected from the group consisting of hydrogen, unsubstituted lower alkyl, xe2x80x94C(O)OR26, and xe2x80x94C(O)NR28R29 where R26 is hydrogen or unsubstituted lower alkyl and R28 and R29 are independently selected from the group consisting of hydrogen or unsubstituted lower alkyl or, combined, R28 and R29 form a group selected from the group consisting of xe2x80x94(CH2)2N(R30)(CH2)2xe2x80x94,
xe2x80x94(CH2)2O(CH2)2xe2x80x94 or xe2x80x94(CH2)5xe2x80x94, R30 being selected from the group consisting of hydrogen and unsubstituted lower alkyl. Preferably R23 is hydrogen, methyl, ethyl, carboxy, ethoxycarbonyl, pyridin-1-ylcarbonyl, piperazin-1-ylcarbonyl, or 4-methylpiperazin-1-ylcarbonyl; or
R23 together with R24 combines to form xe2x80x94(CH2)4xe2x80x94 and xe2x80x94CHxe2x95x90CHxe2x80x94CR34xe2x95x90CHxe2x80x94 R34 is selected from the group consisting of hydrogen and xe2x80x94O(CH2)2NR28R29 and R28 and R29 are independently selected from the group consisting of hydrogen or unsubstituted lower alkyl or, combined, R28 and R29 form a group selected from the group consisting of xe2x80x94(CH2)2N(R30)(CH2)2xe2x80x94, xe2x80x94(CH2)2O(CH2)2xe2x80x94 or xe2x80x94(CH2)5xe2x80x94, R30 being selected from the group consisting of hydrogen and unsubstituted lower alkyl, preferably hydrogen or methyl.
(v) Another preferred group of compounds is that wherein R24 and R25 are independently selected from the group consisting of hydrogen, unsubstituted lower alkyl, aryl optionally substituted with a group selected from the group consisting of halo, unsubstituted lower alkoxy, morpholino and 4-formylpiperidinyl,
xe2x80x94(CH2)nC(O)NR28R29, xe2x80x94(CH2)nC(O)OR26, xe2x80x94(CH2) NR28R29, xe2x80x94(CH2)nOR2, xe2x80x94C(O)NH(CH2)nNR28R29, xe2x80x94O(CH2)nNR28R29xe2x80x94O(CH2)nOR26 or, combined, a group selected from the group consisting of xe2x80x94(CH2)2OC(O)xe2x80x94, xe2x80x94(CH2)2N(R30)C(O)xe2x80x94, xe2x80x94(CH2)5xe2x80x94, xe2x80x94CHxe2x95x90CHxe2x80x94CHxe2x95x90CHxe2x80x94 where n is 0 to 3, R26 is selected from the group consisting of hydrogen and unsubstituted lower alkyl and R28 and R29 are independently selected from the group consisting of hydrogen, unsubstituted lower alkyl, lower alkyl substituted with a phenyl or pyridyl group or xe2x80x94NRR where each R is independently hydrogen or unsubstituted lower alkyl; or R28 and R29 combine to form a group selected from the group consisting of xe2x80x94(CH2)5xe2x80x94, xe2x80x94(CH2)2NR30(CH2)2xe2x80x94 and xe2x80x94(CH2)2O(CH2)2xe2x80x94 where R30 is selected from the group consisting of hydrogen, unsubstituted lower alkyl and xe2x80x94C(O)R26.
Preferably Q is 3,5-dimethyl-4-(4-methylpiperazin-1-yl-carbonyl)-1H-pyrrol-2-yl, 5-(methyl-3H-imidazol-4-yl)-1H-pyrrol-2-yl, 3-methyl-4-(4-methylpiperidin-1-yl-carbonyl)-1H-pyrrol-2-yl, 3,5-dimethyl-1H-pyrrol-2-yl, 3-(2-carboxyethyl)-4,5,6,7-tetrahydro-1H-indol-2-yl, 3-(2-carboxyethyl)-5-methyl-1H-pyrrol-2-yl, 3-(2-carboxyethyl)-5-ethyl-1H-pyrrol-2-yl, 3-(2-carboxyethyl)-4-ethoxycarbonyl-5-methyl-1H-pyrrol-2-yl, 4-(2-carboxyethyl)-3,5-dimethyl-1H-pyrrol-2-yl, 4-(carboxymethyl)-3,5-dimethyl-1H-pyrrol-2-yl, indol-2-yl, 4,5,6,7-tetrahydroindol-2-yl, 5-(2-morpholin-4-ylethyloxy)indol-2-yl, 3-(carboxy)-5-methyl-1H-pyrrol-2-yl, 5-carboxy-3-methyl-1H-pyrrol-2-yl, 3-(3-morpholin-4-ylpropyl)-4,5,6,7-tetrahydroindol-2-yl, 4-(2-diethylaminoethylaminocarbonyl)-3,5-dimethyl-1H-pyrrol-2-yl, 4-(4-methylpiperazin-1-ylcarbonyl)-3,5-dimethyl-1H-pyrrol-2-yl, 5-(4-methylpiperazin-1-ylcarbonyl)-3-methyl-1H-pyrrol-2-yl, 5-(ethoxycarbonyl)-4,5,6,7-tetrahydro-2H-isoindol-3-yl, 4-(pyridin-4-ylaminocarbonyl)-3-phenyl-5-methyl-1H-pyrrol-2-yl, 5-methylthiophen-2-yl, 3-(2-carboxyethyl)-5-ethoxycarbonyl-4-methyl-1H-pyrrol-2-yl, 3-(2-carboxyethyl)-4-carboxy-1H-pyrrol-2-yl, 3-(4-hydroxyphenyl)-4-ethoxycarbonyl-1H-pyrrol-2-yl, 4-(morpholin-4-ylcarbonyl)-3-methyl-1H-pyrrol-2-yl, 4-(piperidin-1-ylcarbonyl)-3-methyl-1H-pyrrol-2-yl, 3-(2-carboxyethyl)-5-(ethoxycarbonyl)-4-methyl-1H-pyrrol-2-yl, 3-(2-carboxyethyl)-4-(carboxy)-1H-pyrrol-2-yl, 3-(methyl)-4-(benzylaminocarbonyl)-1H-pyrrol-2-yl, 3-methyl-4-(pyridin-4-ylmethylaminocarbonyl)-1H-pyrrol-2-yl, 3-methyl-4-[3-(2-oxopyrrolidin-1-yl)propyl-aminocarbonyl)-1H-pyrrol-2-yl, 5-methyl-4-ethoxycarbonyl-3-[3-(4-methylpiperazin-1-yl)propyl]-1H-pyrrol-2-yl, or 3,5-dimethyl-4-(4-methylpiperazin-1-ylaminocarbonyl)-1H-pyrrol-2-yl.
(vi) Yet another preferred group of compounds is that wherein Q is selected from the group consisting of: 
In the above groups (i-vi), a more preferred group of compounds is that wherein R1 and R2 are hydrogen.
(vii) Another preferred group of compounds is represented by the formula (Ia): 
wherein:
Het is 2-, 3-, or 4-pyridyl, pyrimidin-5-yl, thiazol-2-yl, or 2-, 3-, or 4-piperidinyl; and
Q is either:
(a) 
wherein:
J1 is nitrogen and J2, J3 and J4 are carbon and other groups are those defined in the Summary of the Invention.
Within this group a more preferred group of compounds is that wherein R22 is hydrogen.
Within the more preferred group, an even more preferred group of compounds is that wherein:
R23 is selected from the group consisting of hydrogen, unsubstituted lower alkyl, xe2x80x94C(O)OR26, and xe2x80x94C(O)NR28R29 where R26 is hydrogen or unsubstituted lower alkyl and R28 and R29 are independently selected from the group consisting of hydrogen or unsubstituted lower alkyl or, combined, R28 and R29 form a group selected from the group consisting of xe2x80x94(CH2)2N(R30)(CH2)2xe2x80x94,
xe2x80x94(CH2)2O(CH2)2xe2x80x94 or xe2x80x94(CH2)5xe2x80x94, R30 being selected from the group consisting of hydrogen and unsubstituted lower alkyl. Preferably R23 is hydrogen, methyl, ethyl, carboxy, ethoxycarbonyl, pyridin-1-ylcarbonyl, piperazin-1-ylcarbonyl, or 4-methylpiperazin-1-ylcarbonyl; or R23 together with R24 combines to form xe2x80x94(CH2)4xe2x80x94 and xe2x80x94CHxe2x95x90CHxe2x80x94CR34xe2x95x90CHxe2x80x94 R34 is selected from the group consisting of hydrogen and xe2x80x94O(CH2)2NR28R29 and R28 and R29 are independently selected from the group consisting of hydrogen or unsubstituted lower alkyl or, combined, R28 and R29 form a group selected from the group consisting of xe2x80x94(CH2)2N(R30)(CH2)2xe2x80x94, xe2x80x94(CH2)2O(CH2)2xe2x80x94 or xe2x80x94(CH2)5xe2x80x94, R30 being selected from the group consisting of hydrogen and unsubstituted lower alkyl, preferably hydrogen or methyl.
Another even more preferred group of compounds is that wherein R24 and R25 are independently selected from the group consisting of hydrogen, unsubstituted lower alkyl, aryl optionally substituted with a group selected from the group consisting of halo, unsubstituted lower alkoxy, morpholino and 4-formylpiperidinyl, xe2x80x94(CH2)nC(O)NR28R29,xe2x80x94(CH2)nC(O)OR26, xe2x80x94(CH2)nNR28R29, xe2x80x94(CH2)nOR26, xe2x80x94C(O)NH(CH2)nNR28R29, xe2x80x94O(CH2)nNR28R29, xe2x80x94O(CH2)nOR26 or, combined, a group selected from the group consisting of xe2x80x94(CH2)2OC(O)xe2x80x94, xe2x80x94(CH2)2N(R30)C(O)xe2x80x94, xe2x80x94(CH2)5xe2x80x94, xe2x80x94CHxe2x95x90CHxe2x80x94CHxe2x95x90CHxe2x80x94 where n is 0 to 3, R26 is selected from the group consisting of hydrogen and unsubstituted lower alkyl and R28 and R29 are independently selected from the group consisting of hydrogen, unsubstituted lower alkyl, lower alkyl substituted with a phenyl or pyridyl group or xe2x80x94NRR where each R is independently hydrogen or unsubstituted lower alkyl; or R28 and R29 combine to form a group selected from the group consisting of xe2x80x94(CH2)5xe2x80x94, xe2x80x94(CH2)2NR30 (CH2)2xe2x80x94 and xe2x80x94(CH2)2O(CH2)2xe2x80x94 where R30 is selected from the group consisting of hydrogen, unsubstituted lower alkyl and xe2x80x94C(O)R26.
Preferably Q is 3,5-dimethyl-4-(4-methylpiperazin-1-yl-carbonyl)-1H-pyrrol-2-yl, 5-(methyl-3H-imidazol-4-yl)-1H-pyrrol-2-yl, 3-methyl-4-(4-methylpiperidin-1-yl-carbonyl)-1H-pyrrol-2-yl, 3,5-dimethyl-1H-pyrrol-2-yl, 3-(2-carboxyethyl)-4,5,6,7-tetrahydro-1H-indol-2-yl, 3-(2-carboxyethyl)-5-methyl-1H-pyrrol-2-yl, 3-(2-carboxyethyl)-5-ethyl-1H-pyrrol-2-yl, 3-(2-carboxyethyl)-4-ethoxycarbonyl-5-methyl-1H-pyrrol-2-yl, 4-(2-carboxyethyl)-3,5-dimethyl-1H-pyrrol-2-yl, 4-(carboxymethyl)-3,5-dimethyl-1H-pyrrol-2-yl, indol-2-yl, 4,5,6,7-tetrahydroindol-2-yl, 5-(2-morpholin-4-ylethyloxy)indol-2-yl, 3-(carboxy)-5-methyl-1H-pyrrol-2-yl, 5-carboxy-3-methyl-1H-pyrrol-2-yl, 3-(3-morpholin-4-ylpropyl)-4,5,6,7-tetrahydroindol-2-yl, 4-(2-diethylaminoethylaminocarbonyl)-3,5-dimethyl-1H-pyrrol-2-yl, 4-(4-methylpiperazin-1-ylcarbonyl)-3,5-dimethyl-1H-pyrrol-2-yl, 5-(4-methylpiperazin-1-ylcarbonyl)-3-methyl-1H-pyrrol-2-yl, 5-(ethoxycarbonyl)-4,5,6,7-tetrahydro-2H-isoindol-3-yl, 4-(pyridin-4-ylaminocarbonyl)-3-phenyl-5-methyl-1H-pyrrol-2-yl, 5-methylthiophen-2-yl, 3-(2-carboxyethyl)-5-ethoxycarbonyl-4-methyl-1H-pyrrol-2-yl, 3-(2-carboxyethyl)-4-carboxy-1H-pyrrol-2-yl, 3-(4-hydroxyphenyl)-4-ethoxycarbonyl-1H-pyrrol-2-yl, 4-(morpholin-4-ylcarbonyl)-3-methyl-1H-pyrrol-2-yl, 4-(piperidin-1-ylcarbonyl)-3-methyl-1H-pyrrol-2-yl, 3-(2-carboxyethyl)-5-(ethoxycarbonyl)-4-methyl-1H-pyrrol-2-yl, 3-(2-carboxyethyl)-4-(carboxy)-1H-pyrrol-2-yl, 3-(methyl)-4-(benzylaminocarbonyl)-1H-pyrrol-2-yl, 3-methyl-4-(pyridin-4-ylmethylaminocarbonyl)-1H-pyrrol-2-yl, 3-methyl-4-[3-(2-oxopyrrolidin-1-yl)propyl-aminocarbonyl)-1H-pyrrol-2-yl, 5-methyl-4-ethoxycarbonyl-3-[3-(4-methylpiperazin-1-yl)propyl]-1H-pyrrol-2-yl, or 3,5-dimethyl-4-(4-methylpiperazin-1-ylaminocarbonyl)-1H-pyrrol-2-yl.
Representative compounds of the invention are disclosed in the Table below:
1. Brief Description of the Tables
TABLE 1 shows the chemical structures of some exemplary compounds of this invention. The compound numbers correspond to the Example numbers in the Examples section. That is, the synthesis of Compound 1 in Table 1 is described in Example 1. The compounds presented in Table 1 are exemplary only and are not to be construed as limiting the scope of this invention in any manner.
TABLE 2 shows the results of biological testing of some exemplary compounds of this invention. The results are reported in terms of IC50, the micromolar (xcexcM) concentration of the compound being tested which causes a 50% change in the activity of the target PK compared to the activity of the PK in a control to which no test compound has been added. Specifically, the results shown indicate the concentration of a test compound needed to cause a 50% reduction of the activity of the target PK. Bioassays which have been or may be used to evaluate compounds are described in detail below.
The PKs whose catalytic activity is modulated by the compounds of this invention include protein tyrosine kinases of which there are two types, receptor tyrosine kinases (RTKs) and cellular tyrosine kinases (CTKs), and serine-threonine kinases (STKs). RTK mediated signal transduction is initiated by extracellular interaction with a specific growth factor (ligand), followed by receptor dimerization, transient stimulation of the intrinsic protein tyrosine kinase activity and phosphorylation. Binding sites are thereby created for intracellular signal transduction molecules and lead to the formation of complexes with a spectrum of cytoplasmic signaling molecules that facilitate the appropriate cellular response (e.g., cell division, metabolic effects on the extracellular microenvironment, etc.). See, Schlessinger and Ullrich, 1992, Neuron 9:303-391.
It has been shown that tyrosine phosphorylation sites on growth factor receptors function as high-affinity binding sites for SH2 (src homology) domains of signaling molecules. Fantl et al., 1992, Cell 69:413-423, Songyang et al., 1994, Mol. Cell. Biol. 14:2777-2785), Songyang et al., 1993, Cell 72:767-778, and Koch et al., 1991, Science 252:668-678. Several intracellular substrate proteins that associate with RTKs have been identified. They may be divided into two principal groups: (1) substrates that have a catalytic domain, and (2) substrates which lack such domain but which serve as adapters and associate with catalytically active molecules. Songyang et al., 1993, Cell 72:767-778. The specificity of the interactions between receptors and SH2 domains of their substrates is determined by the amino acid residues immediately surrounding the phosphorylated tyrosine residue. Differences in the binding affinities between SH2 domains and the amino acid sequences surrounding the phosphotyrosine residues on particular receptors are consistent with the observed differences in their substrate phosphorylation profiles. Songyang et al., 1993, Cell 72:767-778. These observations suggest that the function of each RTK is determined not only by its pattern of expression and ligand availability but also by the array of downstream signal transduction pathways that are activated by a particular receptor. Thus, phosphorylation provides an important regulatory step which determines the selectivity of signaling pathways recruited by specific growth factor receptors, as well as differentiation factor receptors.
STKs, being primarily cytosolic, affect the internal biochemistry of the cell, often as a down-line response to a PTK event. STKs have been implicated in the signaling process which initiates DNA synthesis and subsequent mitosis leading to cell proliferation.
A group of STKs that comprise a particularly attractive therapeutic target for cell proliferative disorders are the cyclin dependent kinases or CDKs. CDKs play a prominent role in control of cellular proliferation. That is, the proliferation of all eukaryotic cells occurs through a continuum of events called the xe2x80x9ccell cycle.xe2x80x9d While in fact a continuum, for purposes of discussion, the cell cycle is conveniently broken down into four phases, G1, S, G2 and M. There is another phase, known as G0, which is not part of the cell cycle per se but rather is a quiescent state in which a cell resides prior to entering the cell cycle at G1. In G1, cellular activity is heavily dependent on the stimulating influence of external growth factors. It is during G1 that the machinery necessary for DNA replication is assembled. Between G1 and S is a critical point called the xe2x80x9crestrictionxe2x80x9d point. At the restriction point a cell must decide whether it is prepared to continue with the cell cycle. If so, the cell commits to entry into S phase at which time it no longer requires the stimulation of external growth factors. Progress through the cell cycle is entirely intracellular from this point. It is in the S phase that DNA is replicated. At the end of S phase and entry into G2, a cell has 4N DNA content. In G2, a cell begins preparation for M phase and cytokinesis. Progression through the cell cycle is regulated by CDKs. As the name suggests, in order to perform their functions, the CDKs require association with cyclin regulatory subunits. Presently, about nine CDKs and about 12 families of cyclins with which the CDKs can interact are known. Two or these, cyclin D/CDK4 and cyclin E/CDK2 are responsible for controlling entry of a cell into G1 from G0, passage of the cell through the restriction point and commitment to S phase. Progress through S phase is driven by cyclin E/CDK2 and cyclin A/CDK2, the latter of which promotes completion of S phase and entry into G2. Finally, progression through G2, DNA segregation and eventual separation of the parent cell into two daughter cells during M phase and subsequent cytokinesis is controlled by cyclins A and B in conjunction with CDK1. Throughout the cell cycle there are checkpoints at which a cell monitors both its external and internal environments to assure that continued progress through the cycle is appropriate. Two important, well-studied check points occur in G1 and G2/mitosis. At the G1 checkpoint, the cell checks to see that it has adequate nutrition, that it is properly interacting with other cells or their substratum and that its DNA is intact. At the G2 checkpoint, the cell assures that DNA replication is complete and correct and that the mitotic spindle has properly formed. A negative response at any of these checkpoints results in arrest of the cell cycle which can be temporary, if repairs can be made, or permanent, that is, death of the cell, if repairs cannot be made. These checkpoints are important because inappropriate cell cycle progress is a hallmark of cell proliferation disorders such as malignant tumor growth. Since CDKs are primarily responsible for driving cells through the cell cycle, including the checkpoints, their proper functioning is critical to proper cell proliferation. It is for this reason that CDKs have attracted much interest as therapeutic targets. While therapeutic potential exists in all the CDKs, CDK2 has come under particular scrutiny due to the apparently critical role that it play in the cell cycle. For example, it has been demonstrated that CDK2 dominant negative constructs can halt cell cycle progression completely (S. Van den Heuval, et al., Science, 1993, 262:2050-2054). Furthermore, anchorage-independent growth, a key feature of tumor cells, is mediated by CDK2 complexes (G. Orend, et al., Oncogene, 1998, 16:2575-2583). In another study, a peptide inhibitor of CDK2 function was shown to selectively kill tumor cells over normal cells (Y. N. Chen, et al., Proc. Natl. Acad. Sci. USA, 1999, 96:4221-4223).
While not being bound to any theory, a possible mechanism by which a compound capable of mediating CDK2 function might act can be deduced from the relationship of CDK2 and the tumor suppression gene p53. If DNA to be replicated has been damaged or if the cell is being stimulated by an oncogene, p53 is activated by the cell and expresses a protein which either suppresses further cell division or simply instructs the cell to kill itself (apoptosis). However, CDK-2 inhibits the activity of p53, thereby keeping it from performing this crucial function and stimulating cell growth. To counter this, p53 protein stimulates the production of another protein, p21, which complexes with CDK2, thereby inactivating it. However, when the p53 gene is damaged (e.g., mutated, a condition found in most tumor types), the p53-p21/CDK2 complex cell/division-inhibition cascade cannot occur and CDK will stimulate the cell, even though damaged, to divide. This can lead to uncontrolled cellular proliferation and cancer. An exogenous CDK2 inhibitor could, in essence, take the place of p53 and prevent the formation of a cancerous tumor. Thus, one aspect of this invention is a compound which inhibits CDK2 function and thereby the formation of malignant tumors.
Thus it can be seen that PK signal transduction results in, among other responses, cell proliferation, differentiation, growth and metabolism. Abnormal cell proliferation may result in a wide array of disorders and diseases, including the development of neoplasia such as carcinoma, sarcoma, glioblastoma and hemangioma, disorders such as leukemia, psoriasis, arteriosclerosis, arthritis and diabetic retinopathy and other disorders related to uncontrolled angiogenesis and/or vasculogenesis.
A precise understanding of the mechanism by which the compounds of this invention inhibit PKs is not required in order to practice the present invention. However, while not hereby being bound to any particular mechanism or theory, it is believed that the compounds interact with the amino acids in the catalytic region of PKs. PKs typically possess a bi-lobate structure wherein ATP appears to bind in the cleft between the two lobes in a region where the amino acids are conserved among PKs. Inhibitors of PKs are believed to bind by non-covalent interactions such as hydrogen bonding, van der Waals forces and ionic interactions in the same general region where the aforesaid ATP binds to the PKs. More specifically, it is thought that the 2-indolinone component of the compounds of this invention binds in the region normally occupied by the adenine ring of ATP. Specificity of a particular molecule for a particular PK may then arise as the result of additional interactions between the various substituents on the 2-indolinone core and the amino acid domains specific to particular PKs. Thus, different indolinone substituents may contribute to preferential binding to particular PKs. The ability to select compounds capable of binding to the ATP (or other nucleotide) binding site makes the compounds of this invention useful for targeting any protein with such a site. The compounds disclosed herein may thus have utility as in vitro assays for such proteins as well as exhibiting in vivo therapeutic effects through interaction with such proteins.
In another aspect, the protein kinase, the catalytic activity of which is modulated by contact with a compound of this invention, is a protein tyrosine kinase, more particularly, a receptor protein tyrosine kinase. Among the receptor protein tyrosine kinases whose catalytic activity can be modulated with a compound of this invention, or salt thereof, are, without limitation, EGF, HER2,HER3,HER4, IR, IGF-1R, IRR, PDGFRxcex1, PDGFRxcex2, CSFIR, C-Kit, C-fms, Flk-1R, Flk4, KDR/Flk-1, Flt-1, FGFR-1R, FGFR-2R, FGFR-3R and FGFR-4R.
The protein tyrosine kinase whose catalytic activity is modulated by contact with a compound of this invention, or a salt or a prodrug thereof, can also be a non-receptor or cellular protein tyrosine kinase (CTK). Thus, the catalytic activity of CTKs such as, without limitation, Src, Frk, Btk, Csk, Abl, ZAP70, Fes, Fps, Fak, Jak, Ack, Yes, Fyn, Lyn, Lck, Blk, Hck, Fgr and Yrk, may be modulated by contact with a compound or salt of this invention.
Still another group of PKs which may have their catalytic activity modulated by contact with a compound of this invention are the serine-threonine protein kinases such as, without limitation, CDK2 and Raf.
This invention is therefore directed to compounds that modulate PK signal transduction by affecting the enzymatic activity of RTKs, CTKs and/or STKs, thereby interfering with the signals transduced by such proteins. More particularly, the present invention is directed to compounds which modulate RTK, CTK and/or STK mediated signal transduction pathways as a therapeutic approach to the treatment of many kinds of solid tumors, including but not limited to carcinomas, sarcomas including Kaposi""s sarcoma, erythroblastoma, glioblastoma, meningioma, astrocytoma, melanoma and myoblastoma. Treatment or prevention of non-solid tumor cancers such as leukemia are also contemplated by this invention. Indications may include, but are not limited to brain cancers, bladder cancers, ovarian cancers, gastric cancers, pancreas cancers, colon cancers, blood cancers, lung cancers and bone cancers.
Further examples, without limitation, of the types of disorders related to inappropriate PK activity that the compounds described herein may be useful in preventing, treating and studying, are cell proliferative disorders, fibrotic disorders and metabolic disorders.
Cell proliferative disorders, which may be prevented, treated or further studied by the present invention include cancer, blood vessel proliferative disorders and mesangial cell proliferative disorders.
Blood vessel proliferative disorders refer to disorders related to abnormal vasculogenesis (blood vessel formation) and angiogenesis (spreading of blood vessels). While vasculogenesis and angiogenesis play important roles in a variety of normal physiological processes such as embryonic development, corpus luteum formation, wound healing and organ regeneration, they also play a pivotal role in cancer development where they result in the formation of new capillaries needed to keep a tumor alive. Other examples of blood vessel proliferation disorders include arthritis, where new capillary blood vessels invade the joint and destroy cartilage, and ocular diseases, like diabetic retinopathy, where new capillaries in the retina invade the vitreous, bleed and cause blindness.
Two structurally related RTKs have been identified to bind VEGF with high affinity: the fms-like tyrosine 1 (fit-l) receptor (Shibuya et al., 1990, Oncogene, 5:519-524; De Vries et al., 1992, Science, 255:989-991) and the KDR/FLK-1 receptor, also known as VEGF-R2. Vascular endothelial growth factor (VEGF) has been reported to be an endothelial cell specific mitogen with in vitro endothelial cell growth promoting activity. Ferrara and Henzel, 1989, Biochein. Biophys. Res. Comm., 161:851-858; Vaisman et al., 1990, J. Biol. Chem., 265:19461-19566. Information set forth in U.S. application Ser. Nos. 08/193,829, 08/038,596 and 07/975,750, strongly suggest that VEGF is not only responsible for endothelial cell proliferation, but also is the prime regulator of normal and pathological angiogenesis. See generally, Klagsburn and Soker, 1993, Current Biology, 3(10)699-702;Houck, et al., 1992, J. Biol. Chem., 267:26031-26037.
Normal vasculogenesis and angiogenesis play important roles in a variety of physiological processes such as embryonic development, wound healing, organ regeneration and female reproductive processes such as follicle development in the corpus luteum during ovulation and placental growth after pregnancy. Folkman and Shing, 1992, J. Biological Chem., 267(16):10931-34. Uncontrolled vasculogenesis and/or angiogenesis has been associated with diseases such as diabetes as well as with malignant solid tumors that rely on vascularization for growth. Klagsburn and Soker, 1993, Current Biology, 3(10):699-702; Folkham, 1991, J. Natl. Cancer Inst., 82:4-6; Weidner, et al., 1991, New Engl. J. Med., 324:1-5.
The surmised role of VEGF in endothelial cell proliferation and migration during angiogenesis and vasculogenesis indicates an important role for the KDR/FLK-1 receptor in these processes. Diseases such as diabetes mellitus (Folkman, 198, in XIth Congress of Thrombosis and Haemostasis(Verstraeta, et al., eds.), pp. 583-596, Leuven University Press, Leuven) and arthritis, as well as malignant tumor growth may result from uncontrolled angiogenesis. See e.g., Folkman, 1971, N. Engl. J. Med., 285:1182-1186. The receptors to which VEGF specifically binds are an important and powerful therapeutic target for the regulation and modulation of vasculogenesis and/or angiogenesis and a variety of severe diseases which involve abnormal cellular growth caused by such processes. Plowman, et al., 1994, DNandP, 7(6):334-339. More particularly, the KDR/FLK-1 receptor""s highly specific role in neovascularization make it a choice target for therapeutic approaches to the treatment of cancer and other diseases which involve the uncontrolled formation of blood vessels.
Thus, one aspect of the present invention relates to compounds capable of regulating and/or modulating tyrosine kinase signal transduction including KDR/FLK-1 receptor signal transduction in order to inhibit or promote angiogenesis and/or vasculogenesis, that is, compounds that inhibit, prevent, or interfere with the signal transduced by KDR/FLK-1 when activated by ligands such as VEGF. Although it is believed that the compounds of the present invention act on a receptor or other component along the tyrosine kinase signal transduction pathway, they may also act directly on the tumor cells that result from uncontrolled angiogenesis.
Although the nomenclature of the human and murine counterparts of the generic xe2x80x9cflk-Ixe2x80x9d receptor differ, they are, in many respects, interchangeable. The murine receptor, Flk-1, and its human counterpart, KDR, share a sequence homology of 93.4% within the intracellular domain. Likewise, murine FLK-I binds human VEGF with the same affinity as mouse VEGF, and accordingly, is activated by the ligand derived from either species. Millauer et al., 1993, Cell, 72:835-846; Quinn et al., 1993, Proc. Natl. Acad. Sci. USA, 90:7533-7537. FLK-1 also associates with and subsequently tyrosine phosphorylates human RTK substrates (e.g., PLC-xcex3 or p85) when co-expressed in 293 cells (human embryonal kidney fibroblasts).
Models which rely upon the FLK-1 receptor therefore are directly applicable to understanding the KDR receptor. For example, use of the murine FLK-1 receptor in methods which identify compounds that regulate the murine signal transduction pathway are directly applicable to the identification of compounds which may be used to regulate the human signal transduction pathway, that is, which regulate activity related to the KDR receptor. Thus, chemical compounds identified as inhibitors of KDR/FLK-1 in vitro, can be confirmed in suitable in vivo models. Both in vivo mouse and rat animal models have been demonstrated to be of excellent value for the examination of the clinical potential of agents acting on the KDR/FLK-1 induced signal transduction pathway.
Thus, in one aspect, this invention is directed to compounds that regulate, modulate and/or inhibit vasculogenesis and/or angiogenesis by affecting the enzymatic activity of the KDR/FLK-1 receptor and interfering with the signal transduced by KDR/FLK-1. In another aspect, the present invention is directed to compounds which regulate, modulate and/or inhibit the KDR/FLK-1 mediated signal transduction pathway as a therapeutic approach to the treatment of many kinds of solid tumors including, but not limited to, glioblastoma, melanoma and Kaposi""s sarcoma, and ovarian, lung, mammary, prostate, pancreatic, colon and epidermoid carcinoma. In addition, data suggest the administration of compounds which inhibit the KDR/Flk-1 mediated signal transduction pathway may also be used in the treatment of hemangioma, restenosis and diabetic retinopathy.
A further aspect of this invention relates to the inhibition of vasculogenesis and angiogenesis by other receptor-mediated pathways, including the pathway comprising the flt-l receptor.
Receptor tyrosine kinase mediated signal transduction is initiated by extracellular interaction with a specific growth factor (ligand), followed by receptor dimerization, transient stimulation of the intrinsic protein tyrosine kinase activity and autophosphorylation. Binding sites are thereby created for intracellular signal transduction molecules which leads to the formation of complexes with a spectrum of cytoplasmic signaling molecules that facilitate the appropriate cellular response, e.g., cell division and metabolic effects to the extracellular microenvironment. See, Schlessinger and Ullrich, 1992, Neuron, 9:1-20.
The close homology of the intracellular regions of KDR/FLK-1 with that of the PDGF-xcex2 receptor (50.3% homology) and/or the related flt-l receptor indicates the induction of overlapping signal transduction pathways. For example, for the PDGF-xcex2 receptor, members of the src family (Twamley et al., 1993, Proc. Natl. Acad. Sci. USA, 90:7696-7700), phosphatidylinositol-3xe2x80x2-kinase (Hu et al., 1992, Mol. Cell. Biol., 12:981-990), phospholipase cxcex3 (Kashishian and Cooper, 1993, Mol. Cell. Biol., 4:49-51), ras-GTPase-activating protein, (Kashishian et al., 1992, EMBO J., 11:1373-1382), PTP-ID/syp (Kazlauskas et al., 1993, Proc. Natl. Acad. Sci. USA, 10 90:6939-6943), Grb2 (Arvidsson et al., 1994, Mol. Cell. Biol., 14:6715-6726), and the adapter molecules Shc and Nck (Nishimura et al., 1993, Mol. Cell. Biol., 13:6889-6896), have been shown to bind to regions involving different autophosphorylation sites. See generally, Claesson-Welsh, 1994, Prog. Growth Factor Res., 5:37-54. Thus, it is likely that signal transduction pathways activated by KDR/FLK-1 include the ras pathway (Rozakis et al., 1992, Nature, 360:689-692), the PI-3xe2x80x2-kinase, the src-mediated and the plcxcex3-mediated pathways. Each of these pathways may play a critical role in the angiogenic and/or vasculogenic effect of KDR/FLK-1 in endothelial cells. Consequently, a still further aspect of this invention relates to the use of the organic compounds described herein to modulate angiogenesis and vasculogenesis as such processes are controlled by these pathways.
Conversely, disorders related to the shrinkage, contraction or closing of blood vessels, such as restenosis, are also implicated and may be treated or prevented by the methods of this invention.
Fibrotic disorders refer to the abnormal formation of extracellular matrices. Examples of fibrotic disorders include hepatic cirrhosis and mesangial cell proliferative disorders. Hepatic cirrhosis is characterized by the increase in extracellular matrix constituents resulting in the formation of a hepatic scar. An increased extracellular matrix resulting in a hepatic scar can also be caused by a viral infection such as hepatitis. Lipocytes appear to play a major role in hepatic cirrhosis. Other fibrotic disorders implicated include atherosclerosis.
Mesangial cell proliferative disorders refer to disorders brought about by abnormal proliferation of mesangial cells. Mesangial proliferative disorders include various human renal diseases such as glomerulonephritis, diabetic nephropathy and malignant nephrosclerosis as well as such disorders as thrombotic microangiopathy syndromes, transplant rejection, and glomerulopathies. The RTK PDGFR has been implicated in the maintenance of mesangial cell proliferation. Floege et al., 1993, Kidney International 43:47S-54S.
Many cancers are cell proliferative disorders and, as noted previously, PKs have been associated with cell proliferative disorders. Thus, it is not surprising that PKs such as, for example, members of the RTK family have been associated with the development of cancer. Some of these receptors, like EGFR (Tuzi et al., 1991, Br. J. Cancer 63:227-233, Torp et al., 1992, APMIS 100:713-719) HER2/neu (Slamon et al., 1989, Science 244:707-712) and PDGF-R (Kumabe et al., 1992, Oncogene, 7:627-633) are over-expressed in many tumors and/or persistently activated by autocrine loops. In fact, in the most common and severe cancers these receptor over-expressions (Akbasak and Suner-Akbasak et al., 1992, J. Neurol. Sci., 111:119-133, Dickson et al., 1992, Cancer Treatment Res. 61:249-273, Korc et al., 1992, J. Clin. Invest. 90:1352-1360) and autocrine loops (Lee and Donoghue, 1992, J. Cell. Biol., 118:1057-1070, Korc et al., supra, Akbasak and Suner-Akbasak et al., supra) have been demonstrated. For example, EGFR has been associated with squamous cell carcinoma, astrocytoma, glioblastoma, head and neck cancer, lung cancer and bladder cancer. HER2 has been associated with breast, ovarian, gastric, lung, pancreas and bladder cancer. PDGFR has been associated with glioblastoma and melanoma as well as lung, ovarian and prostate cancer. The RTK c-met has also been associated with malignant tumor formation. For example, c-met has been associated with, among other cancers, colorectal, thyroid, pancreatic, gastric and hepatocellular carcinomas and lymphomas. Additionally c-met has been linked to leukemia. Over-expression of the c-met gene has also been detected in patients with Hodgkins disease and Burkitts disease.
IGF-IR, in addition to being implicated in nutritional support and in type-II diabetes, has also been associated with several types of cancers. For example, IGF-I has been implicated as an autocrine growth stimulator for several tumor types, e.g. human breast cancer carcinoma cells (Arteaga et al., 1989, J. Clin. Invest. 84:1418-1423) and small lung tumor cells (Macauley et al., 1990, Cancer Res., 50:2511-2517). In addition, IGF-I, while integrally involved in the normal growth and differentiation of the nervous system, also appears to be an autocrine stimulator of human gliomas. Sandberg-Nordqvist et al., 1993, Cancer Res. 53:2475-2478. The importance of IGF-IR and its ligands in cell proliferation is further supported by the fact that many cell types in culture (fibroblasts, epithelial cells, smooth muscle cells, T-lymphocytes, myeloid cells, chondrocytes and osteoblasts (the stem cells of the bone marrow)) are stimulated to grow by IGF-I. Goldring and Goldring, 1991, Eukaryotic Gene Expression, 1:301-326. Baserga and Coppola suggest that IGF-IR plays a central role in the mechanism of transformation and, as such, could be a preferred target for therapeutic interventions for a broad spectrum of human malignancies. Baserga, 1995, Cancer Res., 55:249-252, Baserga, 1994, Cell 79:927-930, Coppola et al., 1994, Mol. Cell. Biol., 14:4588-4595.
STKs have been implicated in many types of cancer including, notably, breast cancer (Cance, et al., Int. J. Cancer, 54:571-77 (1993)).
The association between abnormal PK activity and disease is not restricted to cancer. For example, RTKs have been associated with diseases such as psoriasis, diabetes mellitus, endometriosis, angiogenesis, atheromatous plaque development, Alzheimer""s disease, restenosis, von Hippel-Lindau disease, epidermal hyperproliferation, neurodegenerative diseases, age-related macular degeneration and hemangiomas. For example, EGFR has been indicated in corneal and dermal wound healing. Defects in Insulin-R and IGF-1R are indicated in type-II diabetes mellitus. A more complete correlation between specific RTKs and their therapeutic indications is set forth in Plowman et al., 1994, DNandP 7:334-339.
As noted previously, not only RTKs but CTKs including, but not limited to, src, abl, fps, yes, fyn, lyn, lck, blk, hck, fgr and yrk (reviewed by Bolen et al., 1992, FASEB J., 6:3403-3409) are involved in the proliferative and metabolic signal transduction pathway and thus could be expected, and have been shown, to be involved in many PTK-mediated disorders to which the present invention is directed. For example, mutated src (v-src) has been shown to be an oncoprotein (pp60v-src) in chicken. Moreover, its cellular homolog, the proto-oncogene pp60c-src transmits oncogenic signals of many receptors. Over-expression of EGFR or HER2/neu in tumors leads to the constitutive activation of pp60cxe2x96xa1src, which is characteristic of malignant cells but absent in normal cells. On the other hand, mice deficient in the expression of c-src exhibit an osteopetrotic phenotype, indicating a key participation of c-src in osteoclast function and a possible involvement in related disorders.
Similarly, Zap70 has been implicated in T-cell signaling which may relate to autoimmune disorders.
STKs have been associated with inflammation, autoimmune disease, immunoresponses, and hyperproliferation disorders such as restenosis, fibrosis, psoriasis, osteoarthritis and rheumatoid arthritis.
PKs have also been implicated in embryo implantation. Thus, the compounds of this invention may provide an effective method of preventing such embryo implantation and thereby be useful as birth control agents.
Finally, both RTKs and CTKs are currently suspected as being involved in hyperimmune disorders.
A method for identifying a chemical compound that modulates the catalytic activity of one or more of the above discussed protein kinases is another aspect of this invention. The method involves contacting cells expressing the desired protein kinase with a compound of this invention (or its salt or prodrug) and monitoring the cells for any effect that the compound has on them. The effect may be any observable, either to the naked eye or through the use of instrumentation, change or absence of change in a cell phenotype. The change or absence of change in the cell phenotype monitored may be, for example, without limitation, a change or absence of change in the catalytic activity of the protein kinase in the cells or a change or absence of change in the interaction of the protein kinase with a natural binding partner.
Examples of the effect of a number of exemplary compounds of this invention on several PKs are shown in Table 2. The compounds and data presented are not to be construed as limiting the scope of this invention in any manner whatsoever.
A compound of the present invention, a prodrug thereof or a physiologically acceptable salt of either the compound or its prodrug, can be administered as such to a human patient or can be administered in pharmaceutical compositions in which the foregoing materials are mixed with suitable carriers or excipient(s). Techniques for formulation and administration of drugs may be found in Remington""s Pharmacological Sciences, Mack Publishing Co., Easton, Pa., latest edition.
Routes of Administration:
As used herein, xe2x80x9cadministerxe2x80x9d or xe2x80x9cadministrationxe2x80x9d refers to the delivery of a compound, salt or prodrug of the present invention or of a pharmaceutical composition containing a compound, salt or prodrug of this invention to an organism for the purpose of prevention or treatment of a PK-related disorder.
Suitable routes of administration may include, without limitation, oral, rectal, transmucosal or intestinal administration or intramuscular, subcutaneous, intramedullary, intrathecal, direct intraventricular, intravenous, intravitreal, intraperitoneal, intranasal, or intraocular injections. The preferred routes of administration are oral and parenteral.
Alternatively, one may administer the compound in a local rather than systemic manner, for example, via injection of the compound directly into a solid tumor, often in a depot or sustained release formulation.
Furthermore, one may administer the drug in a targeted drug delivery system, for example, in a liposome coated with tumor-specific antibody. The liposomes will be targeted to and taken up selectively by the tumor.
Composition/Formulation:
Pharmaceutical compositions of the present invention may be manufactured by processes well known in the art, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or lyophilizing processes.
Pharmaceutical compositions for use in accordance with the present invention may be formulated in conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen.
For injection, the compounds of the invention may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hanks"" solution, Ringer""s solution, or physiological saline buffer. For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.
For oral administration, the compounds can be formulated by combining the active compounds with pharmaceutically acceptable carriers well known in the art. Such carriers enable the compounds of the invention to be formulated as tablets, pills, lozenges, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a patient. Pharmaceutical preparations for oral use can be made using a solid excipient, optionally grinding the resulting mixture, and processing the mixture of granules, after adding other suitable auxiliaries if desired, to obtain tablets or dragee cores. Useful excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol, cellulose preparations such as, for example, maize starch, wheat starch, rice starch and potato starch and other materials such as gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinyl-pyrrolidone (PVP). If desired, disintegrating agents may be added, such as cross-linked polyvinyl pyrrolidone, agar, or alginic acid. A salt such as sodium alginate may also be used.
Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of active compound doses.
Pharmaceutical compositions which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push-fit capsules can contain the active ingredients in admixture with a filler such as lactose, a binder such as starch, and/or a lubricant such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. Stabilizers may be added in these formulations, also.
For administration by inhalation, the compounds for use according to the present invention are conveniently delivered in the form of an aerosol spray using a pressurized pack or a nebulizer and a suitable propellant, e.g., without limitation, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetra-fluoroethane or carbon dioxide. In the case of a pressurized aerosol, the dosage unit may be controlled by providing a valve to deliver a metered amount. Capsules and cartridges of, for example, gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.
The compounds may also be formulated for parenteral administration, e.g., by bolus injection or continuous infusion. Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulating materials such as suspending, stabilizing and/or dispersing agents.
Pharmaceutical compositions for parenteral administration include aqueous solutions of a water soluble form, such as, without limitation, a salt, of the active compound. Additionally, suspensions of the active compounds may be prepared in a lipophilic vehicle. Suitable lipophilic vehicles include fatty oils such as sesame oil, synthetic fatty acid esters such as ethyl oleate and triglycerides, or materials such as liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers and/or agents that increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.
Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g., sterile, pyrogen-free water, before use.
The compounds may also be formulated in rectal compositions such as suppositories or retention enemas, using, e.g., conventional suppository bases such as cocoa butter or other glycerides.
In addition to the formulations described previously, the compounds may also be formulated as depot preparations. Such long acting formulations may be administered by implantation (for example, subcutaneously or intramuscularly) or by intramuscular injection. A compound of this invention may be formulated for this route of administration with suitable polymeric or hydrophobic materials (for instance, in an emulsion with a pharmacologically acceptable oil), with ion exchange resins, or as a sparingly soluble derivative such as, without limitation, a sparingly soluble salt.
A non-limiting example of a pharmaceutical carrier for the hydrophobic compounds of the invention is a cosolvent system comprising benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer and an aqueous phase such as the VPD co-solvent system. VPD is a solution of 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant Polysorbate 80, and 65% w/v polyethylene glycol 300, made up to volume in absolute ethanol. The VPD co-solvent system (VPD:D5W) consists of VPD diluted 1:1 with a 5% dextrose in water solution. This co-solvent system dissolves hydrophobic compounds well, and itself produces low toxicity upon systemic administration. Naturally, the proportions of such a co-solvent system may be varied considerably without destroying its solubility and toxicity characteristics. Furthermore, the identity of the co-solvent components may be varied: for example, other low-toxicity nonpolar surfactants may be used instead of Polysorbate 80, the fraction size of polyethylene glycol may be varied, other biocompatible polymers may replace polyethylene glycol, e.g., polyvinyl pyrrolidone, and other sugars or polysaccharides may substitute for dextrose.
Alternatively, other delivery systems for hydrophobic pharmaceutical compounds may be employed. Liposomes and emulsions are well known examples of delivery vehicles or carriers for hydrophobic drugs. In addition, certain organic solvents such as dimethylsulfoxide also may be employed, although often at the cost of greater toxicity.
Additionally, the compounds may be delivered using a sustained-release system, such as semipermeable matrices of solid hydrophobic polymers containing the therapeutic agent. Various sustained-release materials have been established and are well known by those skilled in the art. Sustained-release capsules may, depending on their chemical nature, release the compounds for a few weeks up to over 100 days. Depending on the chemical nature and the biological stability of the therapeutic reagent, additional strategies for protein stabilization may be employed.
The pharmaceutical compositions herein also may comprise suitable solid or gel phase carriers or excipients. Examples of such carriers or excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycols.
Many of the PK modulating compounds of the invention may be provided as physiologically acceptable salts wherein the claimed compound may form the negatively or the positively charged species. Examples of salts in which the compound forms the positively charged moiety include, without limitation, quaternary ammonium (defined elsewhere herein), salts such as the hydrochloride, sulfate, carbonate, lactate, tartrate, maleate, succinate wherein the nitrogen atom of the quaternary ammonium group is a nitrogen of the selected compound of this invention which has reacted with the appropriate acid. Salts in which a compound of this invention forms the negatively charged species include, without limitation, the sodium, potassium, calcium and magnesium salts formed by the reaction of a carboxylic acid group in the compound with an appropriate base (e.g. sodium hydroxide (NaOH), potassium hydroxide (KOH), Calcium hydroxide (Ca(OH)2), etc.).
Dosage:
Pharmaceutical compositions suitable for use in the present invention include compositions wherein the active ingredients are contained in an amount sufficient to achieve the intended purpose, e.g., the modulation of PK activity or the treatment or prevention of a PK-related disorder.
More specifically, a therapeutically effective amount means an amount of compound effective to prevent, alleviate or ameliorate symptoms of disease or prolong the survival of the subject being treated.
Determination of a therapeutically effective amount is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein.
For any compound used in the methods of the invention, the therapeutically effective amount or dose can be estimated initially from cell culture assays. Then, the dosage can be formulated for use in animal models so as to achieve a circulating concentration range that includes the IC50 as determined in cell culture (i.e., the concentration of the test compound which achieves a half-maximal inhibition of the PK activity). Such information can then be used to more accurately determine useful doses in humans.
Toxicity and therapeutic efficacy of the compounds described herein can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., by determining the IC50 and the LD50 (both of which are discussed elsewhere herein) for a subject compound. The data obtained from these cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage may vary depending upon the dosage form employed and the route of administration utilized. The exact formulation, route of administration and dosage can be chosen by the individual physician in view of the patient""s condition. (See e.g., Fingl, et al., 1975, in xe2x80x9cThe Pharmacological Basis of Therapeuticsxe2x80x9d, Ch. 1 p.1).
Dosage amount and interval may be adjusted individually to provide plasma levels of the active species which are sufficient to maintain the kinase modulating effects. These plasma levels are referred to as minimal effective concentrations (MECs). The MEC will vary for each compound but can be estimated from in vitro data, e.g., the concentration necessary to achieve 50-90% inhibition of a kinase may be ascertained using the assays described herein. Dosages necessary to achieve the MEC will depend on individual characteristics and route of administration. HPLC assays or bioassays can be used to determine plasma concentrations.
Dosage intervals can also be determined using MEC value. Compounds should be administered using a regimen that maintains plasma levels above the MEC for 10-90% of the time, preferably between 30-90% and most preferably between 50-90%.
In cases of local administration or selective uptake, the effective local concentration of the drug may not be related to plasma concentration and other procedures known in the art may be employed to determine the correct dosage amount and interval.
The amount of a composition administered will, of course, be dependent on the subject being treated, the severity of the affliction, the manner of administration, the judgment of the prescribing physician, etc.
Packaging:
The compositions may, if desired, be presented in a pack or dispenser device, such as an FDA approved kit, which may contain one or more unit dosage forms containing the active ingredient. The pack may for example comprise metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser may also be accompanied by a notice associated with the container in a form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the compositions or of human or veterinary administration. Such notice, for example, may be of the labeling approved by the U.S. Food and Drug Administration for prescription drugs or of an approved product insert. Compositions comprising a compound of the invention formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition. Suitable conditions indicated on the label may include treatment of a tumor, inhibition of angiogenesis, treatment of fibrosis, diabetes, and the like.
It is also an aspect of this invention that a compound described herein, or its salt or prodrug, might be combined with other chemotherapeutic agents or cyclooxygenase-2 (COX-2) inhibitors for the treatment of the diseases and disorders discussed above. For instance, a compound, salt or prodrug of this invention might be combined with alkylating agents such as fluorouracil (5-FU) alone or in further combination with leukovorin; or other alkylating agents such as, without limitation, other pyrimidine analogs such as UFT, capecitabine, gemcitabine and cytarabine, the alkyl sulfonates, e.g., busulfan (used in the treatment of chronic granulocytic leukemia), improsulfan and piposulfan; aziridines, e.g., benzodepa, carboquone, meturedepa and uredepa; ethyleneimines and methylmelamines, e.g., altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and trimethylolmelamine; and the nitrogen mustards, e.g., chlorambucil (used in the treatment of chronic lymphocytic leukemia, primary macroglobulinemia and non-Hodgkin""s lymphoma), cyclophosphamide (used in the treatment of Hodgkin""s disease, multiple myeloma, neuroblastoma, breast cancer, ovarian cancer, lung cancer, Wilm""s tumor and rhabdomyosarcoma), estramustine, ifosfamide, novembrichin, prednimustine and uracil mustard (used in the treatment of primary thrombocytosis, non-Hodgkin""s lymphoma, Hodgkin""s disease and ovarian cancer); and triazines, e.g., dacarbazine (used in the treatment of soft tissue sarcoma).
Likewise a compound, salt or prodrug of this invention might be expected to have a beneficial effect in combination with other antimetabolite chemotherapeutic agents such as, without limitation, folic acid analogs, e.g. methotrexate (used in the treatment of acute lymphocytic leukemia, choriocarcinoma, mycosis fungiodes breast cancer, head and neck cancer and osteogenic sarcoma) and pteropterin; and the purine analogs such as mercaptopurine and thioguanine which find use in the treatment of acute granulocytic, acute lymphocytic and chronic granulocytic leukemias.
A compound, salt or prodrug of this invention might also be expected to prove efficacious in combination with natural product based chemotherapeutic agents such as, without limitation, the vinca alkaloids, e.g., vinblastin (used in the treatment of breast and testicular cancer), vincristine and vindesine; the epipodophylotoxins, e.g., etoposide and teniposide, both of which are useful in the treatment of testicular cancer and Kaposi""s sarcoma; the antibiotic chemotherapeutic agents, e.g., daunorubicin, doxorubicin, epirubicin, mitomycin (used to treat stomach, cervix, colon, breast, bladder and pancreatic cancer), dactinomycin, temozolomide, plicamycin, bleomycin (used in the treatment of skin, esophagus and genitourinary tract cancer); and the enzymatic chemotherapeutic agents such as L-asparaginase.
In addition to the above, a compound, salt or prodrug of this invention might be expected to have a beneficial effect used in combination with the platinum coordination complexes (cisplatin, etc.); substituted ureas such as hydroxyurea; methylhydrazine derivatives, e.g., procarbazine; adrenocortical suppressants, e.g., mitotane, aminoglutethimide; and hormone and hormone antagonists such as the adrenocorticosteriods (e.g., prednisone), progestins (e.g., hydroxyprogesterone caproate); estrogens (e.g., diethylstilbesterol); antiestrogens such as tamoxifen; androgens, e.g., testosterone propionate; and aromatase inhibitors such as anastrozole.
The combination of a compound of this invention might be expected to be particularly effective in combination with mitoxantrone or paclitaxel for the treatment of solid tumor cancers or leukemias such as, without limitation, acute myelogenous (non-lymphocytic) leukemia.
Lastly, in addition to the above, a compound, salt or prodrug of this invention might be expected to have a beneficial effect used in combination with COX-2 inhibitor. To treat inflammation. COX-2 inhibitors for use in combination with a compound, salt or prodrug of the preferred embodiments of the present invention might include, without limitation, those disclosed in WO 96/41626 and U.S. Pat. No. 6,248,745. Other COX-2 inhibitors for use in the combinations of the invention include those disclosed in Drugs of the Future, 1997, 22, 711-714 which document is incorporated herein by reference, namely Meloxicam, L-745337 (Merck), MK-966 (Merck), L-768277 (Merck), GR-253035 (Glaxo-Wellcome), JTE-522 (Japan Tobacco), RS-57067-000 (Roche), SC-58125 (Searle), SC-078 (Searle), PD-138387 (Warner-Lambert), NS-398 (Taisho), flosulide and PD-164387 (Warner-Lambert).